Reaction processes such as chemical and biochemical reactions between two or more molecules in vitro, are frequently carried out within a chamber or container. Such chambers, or reaction vessels, are commonly made of glass or plastic and include, for example, test tubes, micro-centrifuge tubes, capillary tubes and microtiter plates.
These processes commonly involve frequent manual handling of samples like, for instance, the transfer of liquids between containers by pipetting, mixing, centrifugation, incubation, washing and precipitation, just to mention a few procedures. Experimental work is accordingly very time consuming and labour intensive.
Additionally, a general characteristic of methods in, for example, molecular biology is that some of the reagents are available only in small amounts or are very expensive, and therefore in most cases only very small volumes, usually in the μl range, of reagents and reaction mixtures are handled in each step of such a method. Reaction chambers currently in use, however, are not amenable for use with volumes below one microliter, due to, e.g., evaporative losses of the reaction solution.
A solution is provided by the miniaturization of those reactions. For example, WO 01/61054 discloses apparatus and methods for parallel processing of micro-volume liquid reactions.
WO 01/12846 in name of PamGene BV relates to the preparation of metal oxide supports loaded with biomolecules.
WO 99/02266 in name of AKZO NOBEL NV relates to a device for performing an assay, a method for manufacturing said device, and use of a membrane in the manufacture of said device.
WO 00/31304 in name of The Perkin Elmer Corporation relates to methods and apparatus for flow-through hybridization.
WO 95/11755 in name of Houston Advanced Research Center relates to microfabricated, flowthrough porous apparatus for discrete detection of binding reactions.
Van Beuningen et al. (2001 Clinical Chemistry 47: 1931-1933) relates to a fast and specific hybridization using flow-through microarrays on porous metal oxide.
However, neither of the above cited prior art documents relates to performing and monitoring successive reactions.
As will be appreciated in the art, there is a continuous need for improved methods and apparatuses suitable for micro-volume reactions. Therefore the present invention aims to provide an integrated high-throughput method for miniaturized molecular reaction processes.